Compositions for enhancing the antibacterial activity of myeloperoxidase and methods of use thereof

Abstract

Methods and compositions are provided for inhibiting the growth of susceptible microorganisms by contacting the microorganisms, in the presence of a peroxide and chloride or bromide, with myeloperoxidase and at least two activity enhancing agent amino acids.

The invention claimed is: 
     
       1. A composition for inhibiting the growth of susceptible microorganisms comprising myeloperoxidase, glucose oxidase, sodium chloride, polysorbate 80, phosphate buffer, and two amino acids, wherein one of the two amino acids is glycine, and the other of the two amino acids is selected from the group consisting of D-isoleucine; L-isoleucine; L-tyrosine; and L-valine; and wherein the two amino acids work in combination to enhance the microbicidal activity of the myeloperoxidase. 
     
     
       2. The composition of  claim 1 , wherein glucose oxidase is present in an amount effective to generate from 100 pmol to 50 μmol peroxide per ml per minute when in the presence of a substrate for the oxidase. 
     
     
       3. The composition of  claim 1  which comprises from 1 to 50,000 μg/ml of myeloperoxidase. 
     
     
       4. The composition of  claim 1  which comprises from 0.1 to about 500 mM of each of the two amino acids. 
     
     
       5. A composition for killing or inhibiting the growth of susceptible microorganisms, wherein the composition consists of myeloperoxidase, glycine,  L -alanine,  L -proline, glucose oxidase, sodium chloride, polysorbate-80, sodium phosphate buffer, and water, wherein the glycine, L-alanine, and L-proline in the composition work in combination to enhance the microbicidal activity of the myeloperoxidase. 
     
     
       6. A method of treating a microbial infection in a human or animal subject, comprising administering to a site of infection in the subject a composition of  claim 1 . 
     
     
       7. The method of  claim 6  wherein the composition comprises glucose oxidase in an amount effective to generate from 100 pmol to 50 μmol peroxide per ml per minute when in the presence of a substrate for the oxidase. 
     
     
       8. The method of  claim 6  wherein the composition comprises from 1 to 50,000 μg/ml of myeloperoxidase. 
     
     
       9. The method of  claim 6  wherein the composition comprises from 0.1 to about 500 mM of each of the two amino acids. 
     
     
       10. The method of  claim 6  wherein the human or animal subject is suffering from a microbial infection of the gums, eyes, ears, skin, soft tissue, wounds, vaginal areas, groin areas, bed sores or burn areas. 
     
     
       11. The method of  claim 10  wherein the infection is a polymicrobial infection. 
     
     
       12. A method for killing or inhibiting the growth of susceptible microorganisms comprising contacting the microorganisms with the composition of  claim 1 . 
     
     
       13. A binary combination, comprising:
 (a) a first composition comprising myeloperoxidase, glucose oxidase, sodium chloride, polysorbate-80, phosphate buffer, water and two amino acids, wherein one of the two amino acids is glycine, and the other of the two amino acids is selected from the group consisting of D-isoleucine; L-isoleucine; L-tyrosine; and L-valine; and wherein the two amino acids work in combination to enhance the microbicidal activity of the myeloperoxidase; and 
 (b) a second composition comprising glucose and water. 
 
     
     
       14. A binary combination for killing or inhibiting the growth of susceptible microorganisms, consisting of:
 (a) a first composition consisting of myeloperoxidase, glycine, L-alanine, L-proline, glucose oxidase, sodium chloride, polysorbate-80, sodium phosphate buffer, and water, wherein the glycine, L-alanine, and L-proline in the composition work in combination to enhance the microbicidal activity of the myeloperoxidase; and 
 (b) a second composition comprising glucose and water. 
 
     
     
       15. A method of treating a microbial infection in a human or animal subject, comprising administering to a site of infection in the subject a composition of  claim 5 . 
     
     
       16. A method for killing or inhibiting the growth of susceptible microorganisms comprising contacting the microorganisms with the composition of  claim 5 . 
     
     
       17. The method of  claim 15  wherein the human or animal subject is suffering from a microbial infection of the gums, eyes, ears, skin, soft tissue, wounds, vaginal areas, groin areas, bed sores or burn areas. 
     
     
       18. The method of  claim 17  wherein the infection is a polymicrobial infection. 
     
     
       19. The composition of  claim 5 , wherein the myeloperoxidase is present in a concentration front bout 10 μg/ml to about 5,000 μg/ml. 
     
     
       20. The composition of  claim 5 , wherein each of the glycine, L-alanine, and L-proline is present in a concentration from about 0.3 mM to about 50 mM. 
     
     
       21. The composition of  claim 5 , wherein the glucose oxidase is present in a concentration from about 1 U/ml to about 500 U/ml.